Park J C, Sung H J, Lee D H, Park Y H, Cho B K, Suh H
Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.
Yonsei Med J. 2000 Oct;41(5):556-62. doi: 10.3349/ymj.2000.41.5.556.
To determine applicability of the cryopreservation procedure for vessel grafts, the viability of endothelial cells (ECs) among the whole cells in three kinds of organs artery, vein, trachea in mongrel dogs was evaluated on the basis of histological analysis. The Griffonia simplicifolia agglutins-fluorescein isothiocyanate (GSA-FITC) and propidium iodide (PI) double staining methods were combined with flow cytometry (FCM), which was able to simultaneously determine the viability of whole cells and ECs from the same tissue, were performed after harvesting, after antibiotic solution treatment, and after cryopreservation and thawing. In most cases, the viability of ECs is lower than that of whole cells from veins and arteries. The viability of whole cells in veins was maintained until the antibiotic solution treatment and then decreased significantly after cryopreservation and thawing, while the ECs began to decrease significantly after the antibiotic solution treatment and more markedly decreased after thawing. The viability of ECs and whole cells from arteries was similar to that of the veins' conditions. The viability of whole cells from the trachea decreased with a similar pattern to that of the ECs from vessels. In consideration of maintaining cell viability among the three kinds of organs, the viability of arteries was better than that of the others. The cells in the trachea demonstrated a lower viability than the vessels. The effect of antibiotic solution treatment on the reduction of cell viability depends on the treatment time and temperature.
为确定血管移植物冷冻保存程序的适用性,基于组织学分析评估了杂种犬三种器官(动脉、静脉、气管)全细胞中内皮细胞(ECs)的活力。将西非单叶豆凝集素 - 异硫氰酸荧光素(GSA - FITC)和碘化丙啶(PI)双重染色方法与流式细胞术(FCM)相结合,该方法能够在收获后、抗生素溶液处理后以及冷冻保存和解冻后,同时测定来自同一组织的全细胞和ECs的活力。在大多数情况下,ECs的活力低于静脉和动脉全细胞的活力。静脉中全细胞的活力在抗生素溶液处理前保持稳定,在冷冻保存和解冻后显著下降,而ECs在抗生素溶液处理后开始显著下降,解冻后下降更明显。动脉中ECs和全细胞的活力与静脉情况相似。气管全细胞的活力下降模式与血管ECs相似。考虑到三种器官中细胞活力的维持情况,动脉的活力优于其他器官。气管中的细胞活力低于血管。抗生素溶液处理对细胞活力降低的影响取决于处理时间和温度。